Back to EveryPatent.com
United States Patent | 5,583,103 |
Ruoslahti ,   et al. | December 10, 1996 |
The present invention provides a method of inhibiting an activity of TGF.beta. comprising contacting the TGF.beta. with a purified decorin. In a specific embodiment, the present invention relates to the ability of decorin, a 40,000 dalton protein that usually carries a glycosaminoglycan chain, to bind TGF.beta.. The invention also provides a novel cell regulatory factor designated MRF. Also provided are methods of identifying, detecting and purifying cell regulatory factors and proteins which bind and affect the activity of cell regulatory factors.
Inventors: | Ruoslahti; Erkki I. (Rancho Santa Fe, CA); Yamaguchi; Yu (San Diego, CA) |
Assignee: | La Jolla Cancer Research Foundation (La Jolla, CA) |
Appl. No.: | 212311 |
Filed: | March 14, 1994 |
Current U.S. Class: | 514/8; 514/2; 530/350; 530/380; 530/399 |
Intern'l Class: | A61K 038/19; C07K 014/495 |
Field of Search: | 530/399,380,350,300 514/8,2 435/7.1 |
Foreign Patent Documents | |||
282317A2 | Sep., 1988 | EP. | |
282317 | Oct., 1988 | EP. |
Socher et al 1987 PNAS 84:8829. Titani et al 1987 PNAS 84:5610. Sporn et al 1986 Science 233:532. Chiefetz et al. "The Transforming Growth Factor .beta. System . . . " Cell vol. 48 pp. 409-415. Border, Wayne A. et al. "Extracellular Matrix and Glomerular Disease." Seminars in Nephrology. 9:307-317 (1989). Andres et al., Membrane-anchored and Soluble Forms of Betaglycan, a Polymorphic Proteoglycan that Binds Transforming Growth Factor-Beta. J. Cell. Biol. 109:3137-3145 (1989). Kanzaki et al., TGF-Beta 1 Binding Protein: A Component of the Large Latent Complex of TGF-Beta1 with Multiple Repeat Sequences. Cell 61:1051-1061 (1990). Cheifetz et al., Heterodimeric Transforming Growth Factor Beta J. Biol. Chem. 263:10783-10789 (1988a). Cheifetz et al., The Transforming Growth Factor-Beta Receptor Type III is a Membrane Proteoglycan. J. Biol. Chem. 263:16984-16991 (1988b). Fisher et al., Deduced Protein Sequence of Bone Small Proteoglycan I (Biglycan) shows Homology with Proteoglycan II (Decorin) and Several Nonconnective Tissue Proteins in a Variety of Species. J. Biol. Chem. 264:4571-4576 (1989). Patthy, L., Detecting Homology of Distantly Related Proteins with Consensus Sequences. J. Mol. Biol. 198:567-577 (1987). Bassols and Massague, Transforming Growth Factor Beta Regulates the Expression and Structure of Extracellular Matrix Chondroitin/ Dermatan Sulfate Proteoglycans. J. Biol. Chem. 263:3039-3045 (1988). Segarini and Seyedin, The High Molecular Weight Receptor to Transforming Growth Factor-Beta Contains Glycosaminoglycan Chains. J. Biol. Chem. 263:8366-8370 (1988). Cheifetz et al., The Transforming Growth Factor-Beta System, a Complex Pattern of Cross-Reactive Ligands and Receptors. Cell 48:409-415 (1987). Ishihara et al., Involvement of Phosphatidylinositol and Insulin in the Coordinate Regulation of Proteoheparan Sulfate Metabolism and Hepatocyte Growth. J. Biol. Chem. 262:4706-4716 (1987). Day et al., Molecular Cloning and Sequence Analysis of the cDNA for Small Proteoglycan II of Bovine Bone. Biochem. J. 248:801-805 (1987). Fritze et al., An Antiproliferative Heparin Sulfate Species Produced by Postconfluent Smooth Muscle Cells. J. Cell Biol. 100:1041-1049 (1985). Krusius and Ruoslahti, Primary Structure of an Extracellular Matrix Proteoglycan Core Protein Deduced from Cloned cDNA. Proc. Natl. Acad. Sci. USA 83:7683-7687 (1986). Castellot et al., Glomerular Endothelial Cells Secrete a Heparinlike Inhibitor and a Peptide Stimulator of Mesangial Cell Proliferation. Am. J. Pathol. 125:493-500 (1986). Castellot et al., Inhibition of Vascular Smooth Muscle Cell Growth by Endothelial Cell-Derived Heparin. J. Biol. Chem. 257:11256-11260 (1982). Vogel et al., Specific Inhibition of Type I and Type II Collagen Fibrillogenesis by the Small Proteoglycan of Tendon. Biochem. J. 223:587-597 (1984). Massague and Like, Cellular Receptors for Type Beta Transforming Growth Factor. J. Biol. Chem. 260:2636-2645 (1985). Brennan, M. et al., Effect of a Proteoglycan Produced by Rat Tumor Cells on Their Adhesion to Fibronectin-Collagen Substrata.sup.1, Cancer Research 43:4302-4307 (1983). Brennan, M. et al., Chondroitin/Dermatan Sulfate Proteoglycan in Human Fetal Membranes, J. of Biol. Chem. 259:13742-13750 (1984). Kresse, H. et al., Glycosaminoglycan-Free Small Proteoglycan Core Protein Secreted by Fibroblasts from a Patient with a Syndrome Resembling Progeroid, Am. J. Hum. Genet. 41, (1987). Pearson, C. et al., The NH.sub.2 -Terminal Amino Acid Sequence of Bovine Skin Proteodermatan Sulfate, The Journal of Biological Chemistry 258:15101-15104 (1983). Ruoslahti, E., Structure and Biology of Proteoglycans, Ann. Rev. Cell Biol. 4:229-255 (1988). Yamaguchi, Y. and Ruoslahti, E., Expression of Human Proteoglycan in Chinese Hamster Ovary Cells Inhibits Cell Proliferation, Nature 336:244-246 (1988). Yamaguchi, Y. et al., Negative Regulation of Transforming Growth Factor-.beta. by the Proteoglycan Decorin, Nature 346:281-284 (1990). Ishiharat, Masayuki "Involvement of Phosphatidylinositol and insulin in the Coordinate Regulation of Proteoheparan Sulfate Metabolism and Hepatocyte Growth." J. Biol. Chem. 262:4708-4716 (1987). Iozzo, Renato V. "Neoplastic Modulation of Extracellular Matrix: Stimulation of Chondroitin Sulfate Proteoglycan and Hyaluronic Acid Synthesis in Co-Cultures of Human Colon Carcinoma and Smooth Muscle Cells." J. Cell. Biochem. 39:355-378 (1989). |
TABLE I ______________________________________ Decorin-Sepharose affinity chromatography of nonlabeled TGF.beta.-1 monitored by growth inhibition assay in Mv1Lu cells. TGF.beta.-1 (ng) Elution Decorin-Sepharose BSA-Sepharose ______________________________________ Flow through & wash 2.7 (2.3%) 82.0 (93.9%) 3 M NaCl 2.2 (1.8%) 1.3 (1.5%) 8 M Urea 116.0 (95.9%) 4.0 (4.6%) ______________________________________